Journal of Lipid Research, Vol. 21, 223-228, February 1980
Copyright © 1980 by Lipid Research, Inc.

Diacyl, alkenyl, and alkyl ether phospholipids in ejaculated, in utero-, and in vitro-incubated porcine spermatozoa

Rhobert W. Evans , Dix E. Weaver , and Eric D. Clegg

Department of Animal Sciences, Purdue University, West Lafayette, IN 47907

The phospholipids of porcine spermatozoa were analyzed by a hydrolytic procedure directly after ejaculation, and after incubation for 120 min in vitro or in ligated uterine segments of females with induced estrus. Total phospholipid content of ejaculated sperm was 65.7 µg lipid P per 10⁹ sperm, of which 41% was alkyl ether and 23% was alkenyl ether glycerophospholipid. All of the ether phospholipids were choline and ethanolamine glycerophospholipids. In order of decreasing amount (% of total phospholipid), the phospholipids were choline and ethanolamine glycerophospholipids (49.9 and 28.2), sphingolipid (10.6), cardiolipin (5.5), phosphatidylinositol (2.3), phosphatidic acid (1.5), phosphatidylserine (1.2), and phosphatidylglycerol (0.8). Phosphorus-containing sphingolipid separated into two components during thin-layer chromatography. Sphingosine was the only long-chain base identified in either band. Major fatty acids in the band with lower R_f were 16:0 (56%), 20:0 (23%), and 18:0 (11%) plus smaller amounts of 14:0, 18:1, and 22:0, while those in the band with higher R_f were 14:0 (30%), 16:0 (45%), and 18:1 (12%) plus smaller amounts of 18:0, 20:0, and 22:0. Choline was the only water-soluble base present in the lower R_f sphingomyelin while ethanolamine was prevalent in the higher R_f component. Incubation of washed spermatozoa in Ca²⁺-free Ringer-fructose at 37°C for 2 hr produced no significant change in the level of any of the phospholipids. Incubation of washed sperm in the uterus for 2 hr, in the presence of oviductal secretions, produced an increase in phosphatidylcholine from 7.2 to 10.2 µg lipid P per 10⁹ sperm.—Evans, R. W., D. E. Weaver, and E. D. Clegg. Diacyl, alkenyl, and alkyl ether phospholipids in ejaculated, in utero-, and in vitro-incubated porcine spermatozoa.

Supplementary key words sperm • sphingolipids • glycerophospholipids

Submitted on April 4, 1979
Accepted on October 18, 1979

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